Development of novel predictive model for drug susceptibility using next-generation cancer imaging that visualizes intratumoral heterogeneity

• Project/Area Number: 18K19482
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 50:Oncology and related fields
• Research Institution: Keio University
• Principal Investigator: Tanaka Nobuyuki 慶應義塾大学, 医学部(信濃町), 講師 (60445244)
• Project Period (FY): 2018-06-29 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000); Fiscal Year 2018: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: 腎細胞癌 / ライトシート顕微鏡 / 膨張顕微鏡法 / 分子標的治療 / 免疫治療 / 腫瘍内不均一性 / イメージング / In situハイブリダイゼーション

Outline of Final Research Achievements

In this study, we use next-generation cancer imaging "light-sheet microscopy/expansion microscopy" to stereoscopically visualize intratumoral inhomogeneity. We have implemented a new 3D imaging pipeline: DIIFCO method as the basis of this technology in actual clinical tissues. Expansion microscopy is also the latest research method to overcome the diffraction limit of optical microscopy. Utilizing a renal cell carcinoma tissue, we proceeded with the development of a protocol for in situ hybridization by the DIIFCO method under tissue swelling. By combining the expansion microscopy and DIIFCO method, RNA localization can be understood by nano-resolution. In addition, heterogeneous lncRNA expression was also revealed to be intracellularly localized in our methods.

Academic Significance and Societal Importance of the Research Achievements

次世代イメージング「ライトシート顕微鏡・膨張顕微鏡法」は、ヒト癌研究への適用自体が革新的であり、国内外に類似研究は存在しない。3次元イメージングの普及に伴い、今後様々な臓器・細胞において遺伝子発現の空間分布や微小脈管の構造的意義が問われていくと考える。世界に先駆ける申請者の解析プラットフォームは、臨床に直結する橋渡し研究(トランスレーショナルリサーチ)であり、本研究が癌イメージング術の3次元化を促す分水嶺となり得る。確立されるイメージング法は、普遍的な社会への波及効果が期待できる。

Research Products

• [Int'l Joint Research] Karolinska Institutet(スウェーデン)
• [Int'l Joint Research] Medical Biochemistry and Biophysics/Karolinska Institutet/Stockholm(スウェーデン)
• [Journal Article] Three-dimensional single-cell imaging for the analysis of RNA and protein expression in intact tumour biopsies (2020)
  • Author(s): Tanaka Nobuyuki、Kanatani Shigeaki、Kaczynska Dagmara、Fukumoto Keishiro、Louhivuori Lauri、Mizutani Tomohiro、Kopper Oded、Kronqvist Pauliina、Robertson Stephanie、Lindh Claes、Kis Lorand、Pronk Robin、Niwa Naoya、Matsumoto Kazuhiro、Oya Mototsugu、... Uhlen Per
  • Journal Title: Nature Biomedical Engineering Volume: 4 Issue: 9 Pages: 875-888
  • DOI: 10.1038/s41551-020-0576-z
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] TNFAIP2 expression induces epithelial-to-mesenchymal transition and confers platinum resistance in urothelial cancer cells. (2019)
  • Author(s): Niwa N, Tanaka N, Hongo H, Miyazaki Y, Takamatsu K, Mizuno R, Kikuchi E, Mikami S, Kosaka T, Oya M.
  • Journal Title: Laboratory Investigation Volume: 99 Issue: 11 Pages: 1702-1713
  • DOI: 10.1038/s41374-019-0285-y
  • Peer Reviewed
• [Journal Article] Volumetric imaging: a potential tool to stage upper tract urothelial carcinoma. (2019)
  • Author(s): Grahn Alexandra、Tanaka Nobuyuki、 Uhlen Per、 Brehmer Marianne
  • Journal Title: World Journal of Urology Volume: 37 Issue: 11 Pages: 2297-2302
  • DOI: 10.1007/s00345-019-02682-1
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Mapping of the three-dimensional lymphatic microvasculature in bladder tumours using light-sheet microscopy. (2018)
  • Author(s): Tanaka N, Kaczynska D, Kanatani S, Sahlgren C, Mitura P, Stepulak A, Miyakawa A, Wiklund P, Uhlen P.
  • Journal Title: British Journal of Cancer Volume: 118 Issue: 7 Pages: 995-999
  • DOI: 10.1038/s41416-018-0016-y
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Improved Pathological Examination of Tumors with 3D Light-Sheet Microscopy. (2018)
  • Author(s): Uhlen P, Tanaka N.
  • Journal Title: Trends in Cancer Volume: 4 Issue: 5 Pages: 337-341
  • DOI: 10.1016/j.trecan.2018.03.003
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Single-cell RNA-seq analysis reveals the platinum resistance gene COX7B and the surrogate marker CD63 (2018)
  • Author(s): Tanaka Nobuyuki、Katayama Shintaro、Reddy Aparna、Nishimura Kaneyasu、Niwa Naoya、Hongo Hiroshi、Ogihara Koichiro、Kosaka Takeo、Mizuno Ryuichi、Kikuchi Eiji、Mikami Shuji、Miyakawa Ayako、Arenas Ernest、Kere Juha、Oya Mototsugu、Uhl?n Per
  • Journal Title: Cancer Medicine Volume: 7 Issue: 12 Pages: 6193-6204
  • DOI: 10.1002/cam4.1828
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Single-cell RNA-seq analysis of intra-tumor heterogeneity dynamics reveals the novel platinum resistance gene COX7B in patients with urinary bladder cancer (2020)
  • Author(s): 田中伸之、丹羽直也、 本郷周、安水洋太、武田利和、松本一宏、森田伸也、小坂威雄、水野隆一、浅沼宏、Per Uhlen、大家基嗣
  • Organizer: 第108回日本泌尿器科学会総会
• [Presentation] 3次元ライトシート顕微鏡による腫瘍内不均一性の可視化：スライドフリーな1細胞レ ベルのタンパク/RNA発現解析 (2019)
  • Author(s): 田中 伸之、三上 修治、小坂 威雄、水野 隆一、大家 基嗣
  • Organizer: 第78回日本癌学会学術総会
• [Presentation] 3次元ライトシート顕微鏡を用いた次世代泌尿器癌イメージング:腫瘍内不均一 性の可視化 (2019)
  • Author(s): 田中 伸之、三上 修治、桝田 司、武田 利和、松本 一宏、森田 伸也、小坂 威雄、浅沼 宏、水野 隆一、大家 基嗣
  • Organizer: 第57回 日本癌治療学会学術集会
• [Presentation] Transcription and protein phenotyping with 3D pathology: Light-sheet microscopy overlooks cellular malignancy of intact tumour volumes (2019)
  • Author(s): Tanaka N, Kanatani S, Kaczynska D, Louhivuori L, Oya M, Miyakawa A, Uhlen P.
  • Organizer: The 34th Annual EAU Congress
• [Presentation] がん微小環境の可視化：透明化技術を利用した次世代癌イメージング (2018)
  • Author(s): 田中伸之、大家基嗣
  • Organizer: 第83回日本泌尿器科学会東部総会
• [Remarks] 慶應義塾大学医学部 泌尿器科学教室ホームページ
  • URL: http://www.keio-urology.jp/
• [Remarks]
  • URL: http://www.keio-urology.jp/
• [Remarks] 慶應義塾大学医学部泌尿器科学教室
  • URL: http://www.keio-urology.jp/